KR20060111790A - Composition comprising the extract of saururus chinensis for the prevention or treatment of asthma or allegic disease - Google Patents

Abstract

A composition for the prevention or treatment of asthma or allergic disease comprising the extract of Saururus chinensis is provided to inhibit emigration of eosinophil and eotaxin gene, and prevent production of inflammatory eicosanoid by repressing activity of 5-LO(5-lipoxygenase). The composition for the prevention or treatment of asthma or allergic disease comprises the extract of Saururus chinensis, wherein the extract of Saururus chinensis is obtained by extracting Saururus chinensis with polar solvent selected from water, methanol and ethanol; the asthma or allergic disease is bronchial asthma, allergic nasitis, allergic asthma or allergic dermatitis; and the amount of Saururus chinensis extract is 0.01-50% based on the total weight of the composition.

Description

Composition comprising the extract of Saururus chinensis for the prevention or treatment of asthma or allegic disease}

FIG. 1 is a diagram showing a decrease in eosinophil count in an ovalbumin / alum-induced asthma model in order to know the anti-asthma ability of in vivo against trichophytium extract.

Figure 2 is a diagram showing the results on the expression of asthma-related genes from lung tissue of the ovalbumin / alum-induced asthma model to know the anti-asthma ability of the three hundred seconds extract,

Figure 3 is a diagram showing the efficacy in the allergy model induced by IgE / anti-IgE to know the anti-allergic capacity of the three hundred seconds extract,

4 is a diagram showing the inhibitory activity of 5-lipoxygenase involved in the synthesis of leukotriene (LT) C ₄ causing bronchial asthma and allergic reactions.

The present invention relates to a composition for the prevention or treatment of asthma or allergic diseases, and more particularly to a composition for the prevention or treatment of asthma or allergic diseases, including three hundred (S aururus chinensis ) extract as an active ingredient.

Allergic diseases are most often caused by mast cells in tissues activated by an antigen-antibody response and by basophil cells in blood and eosinophils (primarily histamine, leukotrienes, prostaglandins, TNFα, cytokines, etc.). Since the use of allergy therapeutic drugs currently used remains symptomatic, there is an urgent need for the development of more fundamental therapeutic drugs.

Key mediators to induce allergic diseases include prostaglandindes, leukotriens, and platelet activators (PAFs). Phospholipase A2 and cyclooxygenase And lipoxygenase to generate from arachidonic acid as a precursor.

On the other hand, drugs recently attracting attention as an allergic agent for treating asthma are drugs having both histamine free inhibition, leukotriene C ₄ production inhibition, and platelet activator inhibitor activity.

On the other hand, three hundred (S aururus chinensis ) is a perennial herb belonging to the tree three hundred and over, the height 0.5 ~ 1m. Underground stem long stretched sideways, leaves alternate, egg-shaped, basal-shaped. 2-3 leaves under the inflorescence are white. In late June to July, water inflorescences face down and fall down. Inflorescences stand up straight, with small white flowers growing slowly from the base to the end. The flower stalk is not on the inflorescence axis, but is attached to the end of a small peduncle of 2 ~ 3㎜ in length. Flowers are bisexual, without cover. There are 6 surgeries facing the carpel, and the base is attached to the carpel. The pistil consists of four carpels, and the bases stick together. 2 seeds per 1 carpel, but 1 seed per carpel. The leaves, flowers, and roots are white, and two or three leaves on the upper part are called three hundred seconds. It lives in low wetlands and is distributed in Korea, Japan, and Southeast Asia. There are two species of genus 300 , of which S. cernuus is distributed in eastern North America.

 It is also called horse riding or red riding horse. It improves blood circulation, releases blood, lowers fever, releases poison, stops cramps, and stops pain, resulting in overworked illness, sore muscles and bone nodes, soreness and bruises. , Arthralgia, pain after surgery, snake bite poison (Bae Gi Hwan, Korean medicinal plant, Kyohaksa, p204, 2000)

Thus, the present invention has been confirmed that the inhibitory effect in the three hundred seconds extract asthma model and allergic animal model, the decrease in eosinophil count in the asthma model is eosinophil eotaxin (Garcia-Zepeda EA, et al., Nature Medicine, 2 (4) ), pp449-456, 1996) and is essential interleukin (IL) -4 gene in the generation of IgE antibodies of allergic reactions main (Arm JP. et al., strongly Advance Immunology, 51, pp323-382, 1992 ) The present invention was completed by finding out that it possesses the effect of inhibiting leukotriene C ₄ production, which is involved in allergy and asthma, at the same time.

It is an object of the present invention to provide a pharmaceutical composition and dietary supplement comprising the three hundred sec extract as an active ingredient effective for asthma or allergic diseases.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of asthma or allergic diseases containing three hundred (S aururus chinensis ) extract as an active ingredient.

The extract includes extracts available for polar solvents such as water, methanol, ethanol and the like, and mixed solvents thereof.

The asthma or allergic diseases include bronchial asthma, allergic rhinitis, allergic asthma or allergic dermatitis and the like.

Hereinafter, a method of obtaining the three hundred sec extract will be described in detail.

The extract of the present invention, trichophytium, about 3 to 10 times the weight (kg), preferably about 3 to 10 times the water (C ₁ ) by cutting the dried outpost, root or ground (leaf, stem) of three hundred seconds, C ₁ To lower alcohols of C ₄ or a mixed solvent thereof, preferably water or methanol, at an extraction temperature of 20 ° C. to 100 ° C., preferably 50 ° C. to 100 ° C., for about 1 to 10 days, preferably about 2 hours. The extract obtained by using cold needle, hot water extraction, ultrasonic extraction, reflux cooling extraction method for 5 hours may be obtained by filtration, concentration under reduced pressure, or drying.

In another aspect, the present invention provides a pharmaceutical composition containing the three hundred seconds extract obtained by the above method as an active ingredient, and is effective in preventing or treating asthma or allergic diseases.

The composition of the present invention can be applied to bronchial asthma, allergic rhinitis, allergic asthma or allergic dermatitis.

The composition of the present invention preferably contains 0.01 to 99.9% of the tritical herb extract, and more preferably 0.1 to 90%. However, the composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.

The composition comprising the tritical extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Compositions comprising extracts according to the invention are formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Carriers, excipients and diluents which may be used in the composition comprising the extract, may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, Gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.

The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection. Pharmaceutical dosage forms of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

The present invention provides a dietary supplement comprising a three hundred seconds extract and a pharmaceutically acceptable food supplement additive exhibiting the effect of preventing asthma or allergic diseases.

The composition comprising the extract of the present invention can be used in various ways, such as drugs, foods and drinks for the prevention of asthma or allergic diseases. Examples of foods to which the extract may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. .

Since the extract of the three hundred seconds of the present invention has almost no toxicity and side effects, it is a drug that can be used safely even for long-term administration for the purpose of prevention.

The extract of the present invention may be added to food or beverages for the purpose of preventing asthma or allergic diseases. At this time, the amount of the extract in the food or beverage is generally added to the health food composition of the present invention to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 30 g based on 100 ml, preferably Can be added in a ratio of 0.3 to 10 g.

In addition to containing the extract as an essential ingredient in the indicated ratio, the health beverage composition of the present invention is not particularly limited in the liquid component and may contain various flavors or natural carbohydrates as additional ingredients, as in general beverages. Examples of the above-mentioned natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like. Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

Example 1. Preparation of three hundred vine extract

1-1. Method of manufacturing hot water extract

Hot water extract can be obtained by heating 1.3 kg of 300 seconds of starch and root purchased from Gyeongdong market in 50 ml of purified water for 15 minutes.

1-2. Method of making methanol extract

The remaining three hundred seconds except for the hot water extract of Example 1-1 was immersed in methanol, and then extracted. The extract was obtained at room temperature, and extracted once more by adding methanol, and the filtrates were mixed. It was. The methanol extract was concentrated under reduced pressure, and dried to obtain 100 g of 300 sec extract.

1-3. Method of manufacturing ethanol extract of 300 seconds

The remaining layer, except for the methanol extract obtained in Example 1-2, was extracted and fractionated three times with ethanol, and the ethanol soluble layers were combined, followed by a vacuum rotary evaporator (Nihon Seiko, VR-205c, Japan). The solvent was evaporated to give ethanol extract.

Experimental Example 1. Measurement of the effect on eosinophil infiltration in an asthma model induced by Ovalbumin

In order to examine the effect on the anti-asthma efficacy of the three hundred seconds extract of the present invention, the following experiment was performed.

That is, mice (Balb / c female rats, 6 weeks old) are administered with a 1: 1 mixed solution of Ovalbumin 100 ug (Sigma) and Alum (Alum) intraperitoneally three times a day 0, 7 days, 14 days. The first 3 days of week 4 were administered orally twice a day, and the last day was administered orally once a day, for a total of 7 oral doses for 4 days. Ovalbumin is aerosolized in the rat's nose. After killing in the morning on the oily day, 1.5 ml of phosphate buffered saline (PBS) solution was directly administered to the bronchus to obtain bronchial lavage. Thereafter, the washing solution was applied to the slide glass with cytospin, and then stained with Diff-Quick (Japan, Japan), and stained from 200 cells using an optical microscope. (See Figure 1). In the present experiment, dexamethasone (dexamethason; 5 mg / kg oral administration to the same person as the 300 sec extract) was used as a positive control.

Experimental Example 2 Measurement of Cytokine and Eotaxin Gene Expression Inhibition Effect from Lung Tissues of Ovalbumin-Induced Asthma Model

2-1. RNA Preparation and RT-Polymerase Chain Reaction (RT-PCR)

Total RNA was isolated from lung tissues of ovalbumin-induced rats and untreated rats using TRIZOL reagent (Invitrogen, USA) according to the manufacturer's instructions. 1 gram of RNA was added to 1 g of each lung tissue and 1 ㎍ of RNA isolated was reverse transcriptase (TAKARA, Japan), 1 mM dNTP and oligo (dT _12-18 ) 0.5 ㎍ / μl. Was reverse transcribed, and then PCR was purified using cDNA to detect eotaxin, interleukin-4 and β-actin gene expression using a thermal cycler (Perkin Elmer Cetus co., USA). Analysis was performed. The reaction was carried out with 2.5 pmol oligo (dT ₂₀ ) primers, 5 Unit Avian Myeloblastosis Virus (AMV) reverse transcriptase ^R (Takara, Japan), Taq DNA polymerase (Takara 1 Unit), 0.2 mM It was performed in 20 μl containing dNTP, reaction buffer (× 10). After initial denaturation at 95 ° C. for 3 minutes, the cycle number was used within index ranges at 94 ° C. for 30 seconds, at 57 ° C. for 30 seconds, and at 72 ° C. for 30 seconds. PCR was electrophoresed in 1.2% agarose gel, stained with ethidium bromide and detected by ultraviolet light.

As a result, it was confirmed that when treated with 100 mg / ㎖ extract, Eotaxin and interleukin-4 mRNA is significantly reduced dose-dependent by the 300 seconds extract (see Figure 2).

The size of the primer and gene product used in the experiment is as follows.

Eotaxin primer set (gene product; 495 b.p),

Forward Primer: CCA AGG ACT TGG CRR CAT GTA G

Reverse Primer: ATT CTG GCT TGG CAT GGT AGC

Interleukin-4 primer collection (gene product; 454 b.p),

Omni-directional primer: ACG GCA CAG AGC TAT TGA TG

Reverse primer: ATG GTG GCT CAG TAC TAC GA

β-actin (gene product; 574 b.p)

Omnidirectional: 5'-CAC CCG CCA CCA GTT CGC CA-3 '

Reverse: 5'-CAG GTC CCG GCC AGC CAG GT-3 '

Experimental Example 3. Anti-allergic inhibitory effect in animal model

In order to know the anti-allergic ability of the in vivo (in vivo) to the extract of Example 1 was carried out. Ten rats per experimental group were tested using passive cutaenous anaphylaxis for rats, a representative type I allergy model, using Sprague-Dawley rats (Katayama S, et. al., Microbiol Immunol 22, pp89-101, 1978).

Monoclonal anti DNP mouse IgE (Sigma, Inc., 100 μl, 1000-fold diluted solution) was administered to Sprague-Dawley rats 48 hours prior to administration of antigen (DNP, Sigma) 1 Three hundred seconds extract (dissolved in 0.5% CMC) was administered 100-400 mg / kg / 100 μl orally once daily for 3 days, at which time 1-10 mg of prednisolone (prednisolone (Sigma)) was used as a positive control. / Kg / 100 μL were administered intraperitoneally, followed by injection of 1 mg DNP-BSA (antigen solution) / 1% Evans blue solution into the tail vein 30 minutes later, The result is shown in FIG.

As shown in Figure 3, the substance of the present invention showed a significant anti-allergic action in the allergy model in vivo.

Experimental Example 4. Analysis of the effect on the production of leukotriene

4-1. Experiment method

The following experiment was carried out to examine the effect of the extract of the three hundred seconds prepared in the above example on leukotriene C ₄ production.

Mouse bone marrow-derived mast cells (BMMC) were derived from BALB / C mice using Murakami et al. (Murakami et al., J. Biol. Chem. 269 , pp22269-22275, 1994). Isolated from bone marrow and cultured in 50% WEHI-3 conditioned medium (containing 10% FCS) containing culture supernatant of WEHI-3 cells (provided by Professor Kudo Ichiro, Pharmacy, Showa University, Japan) It was.

After 3 weeks of culture, mast cells were treated with SCF / LPS mixed stimulant (Sigma) for 30 minutes. The leukotriene C ₄ quantification of the supernatant after cell stimulation was measured by Enzyme linked immunoassay (EIA) using the leukotriene C ₄ assay kit (Cayman). Three hundred seconds extract was pretreated for 15 minutes in advance, and the amount of leukotriene C ₄ produced by adding a stimulant was measured.

4-2. Experiment result

As shown in FIG. 4, the inhibition of leukotriene C ₄ production of the extract inhibited the production of leukotriene C _{4 in} a dose dependent manner, and the concentration (IC ₅₀ ) required for 50% inhibition was 0.28 μg / ml (see FIG. 4).

Formulation Example 1 Preparation of Injection

Three hundred seconds extract of Example 1 ...... 10 mg

Sodium Metabisulfite ........................ 3.0mg

Methylparaben ............... 0.8 mg

Propylparaben ....................... 0.1mg

Sterile Distilled Water for Injection ...

The above ingredients are mixed and made into 2 ml by a conventional method, and then filled into 2 ml ampoules and sterilized to prepare an injection.

Formulation Example 2 Preparation of Tablet

Three hundred seconds extract of Example 1 ............ 200 mg

Lactose ............... 100 Mg

Starch ........................... 100 Mg

Magnesium Stearate .....................

The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.

Formulation Example 3 Preparation of Capsule

Three hundred seconds extract of Example 1 ...... 10 mg

Lactose ......................... 50 Mg

Starch ... 50 Mg

Talc .................. 2mg

Magnesium Stearate ...............

Capsules are prepared by mixing the above ingredients and filling into gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 4 Preparation of Liquid

Three hundred seconds extract of Example 1 ............ 1000 mg

Sugar................................................. .20 g

Isomerized sugar ......................................... 20g

Lemon flavor Quantity

Purified water was added to make a total of 1000 ml. According to the conventional method for preparing a liquid, the above components are mixed, and then filled into a brown bottle and sterilized to prepare a liquid.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, use purpose.

Formulation Example 5 Preparation of Healthy Food

Three hundred seconds extract of Example 1 ............. 1000 mg

Vitamin Blend ...............

Vitamin A Acetate ......................... 70 μg

Vitamin E ........................................ 1.0 mg

Vitamin B1 ......................................... 0.13 mg

Vitamin B2 ...................................... 0.15 mg

Vitamin B6 ......................................... 0.5 mg

Vitamin B12 ......................... 0.2 μg

Vitamin C ......................................................... 10 Mg

Biotin ... 10 μg

Nicotinic Acid Amide ... 1.7 mg

Folic Acid ... ..50 ㎍

Calcium Pantothenate ......................................... 0.5 mg

Mineral mixture ........................

Ferrous Sulfate ............... 1.75 mg

Zinc Oxide ......................................... 0.82 mg

Magnesium Carbonate ......................................... 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dicalcium Phosphate Dibasic ......................................... 55 mg

Potassium Citrate ... 90 mg

Calcium Carbonate ... 100 mg

Magnesium Chloride ......................................... 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 6 Preparation of Healthy Drink

Three hundred seconds extract of Example 1 ... 1000 mg

Citric Acid ........................................ 1000 mg

Oligosaccharide ........................................ 100 g

Plum concentrate ........................................ 2 g

Taurine ... .1 g

Purified water is added to the whole ..... 900 ml

After mixing the above components according to a conventional healthy beverage manufacturing method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is a composition suitable for a preferred beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, use purpose.

As described above, the three hundred seconds extract of the present invention exhibits an anti-asthma, anti-allergic effect, as a composition for the prevention or treatment of asthma or allergic diseases, can be used as a medicinal and health supplement consisting of non-toxic natural medicine. .

Claims (6)

A pharmaceutical composition for the prevention or treatment of asthma or allergic diseases, including the extract of S aururus chinensis as an active ingredient. The pharmaceutical composition according to claim 1, which is an extract soluble in polar solvents such as water, methanol, ethanol and the like and a mixed solvent thereof. The pharmaceutical composition of claim 1, wherein the asthma or allergic disease is bronchial asthma, allergic rhinitis, allergic asthma or allergic dermatitis. The pharmaceutical composition according to any one of claims 1 to 3, comprising 0.01 to 50% of the extract based on the total weight of the composition. A dietary supplement comprising S aururus chinensis extract and a pharmaceutically acceptable food supplement that exhibits a prophylactic or ameliorating effect of asthma or allergic diseases. 6. The dietary supplement of claim 5 in the form of a powder, granule, tablet, capsule or beverage.

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